Matrix metalloproteinases (MMPs) are a family of at least 15 enzymes that degrade the extracellular matrix (ECM) (Borden, P. and Heller, R. Crit. Rev. Eukaryotic Gene Expr. 7: 159-178, 1997). These enzymes have essential roles in modeling and remodeling the ECM in normal physiology and disease pathology. Several of these enzymes have the unique ability to degrade the interstitial collagenase (types I, II, and III), the body's most abundant proteins. MMP-1 is the most ubiquitously expressed interstitial collagenase, thereby assigning it a prominent role in collagen degradation. Overexpression of MMP-1 is associated with several pathological conditions, including the irreversible degradation of cartilage, tendon, and bone in arthritis (Vincenti et al. Crit. Rev. Eukaryotic Gene Expr. 6:391-411, 1996) and the degradation of collagenase I and III in tumor invasion and metastasis (Chambers, A. F. and Matrisian, L. M. J. Nat'l Cancer Inst. 89:1260-1270, 1997; Murray et al. Nat. Med. 2:461-462, 1996). Patients with tumors that express MMP-1 have an overall poorer prognosis than patients with tumors that do not express this protein (Murray et al. Nat. Med. 2:461-462, 1996; Murray et al. J. Pathol. 185:256-261, 1998). This overexpression of MMP-1 has been suggested to be due to the juxtaposition of transcription factor binding sites within the promoter of this gene and to the cooperativity among the factors that bind these sites (Buttice et al. Oncogene 13:2297-2306, 1996; Basuyaux et al. J. Biol. Chem. 272:26188-26195, 1997; Gutman, A. and Waslyk, B. EMBO J. 9:2241-2246, 1990; Benbow, U. and Brinckerhoff, C. E. Matrix Biol. 15:519-526, 1997).
Most normal cells express modest, but detectable, levels of MMP-1 constitutively, and this expression increases substantially in the presence of cytokines or growth factors (Vincenti et al. Crit. Rev. Eukaryotic Gene Expr. 6:391-411, 1996; Rutter et al. J. Cell Biochem. 66:322-336, 1997; Aho et al. Eur. J. Biochem. 247:503-510, 1997; Delany, A. M. and Brinckerhoff, C. E. J. Cell Biochem. 50:400-410, 1992). However, A2058 melanoma cells constitutively express high levels of MMP-1 (Templeton et al. Cancer Res. 50:5431-5437, 1990), making them a useful model for studies on the transcriptional regulating of this gene and for comparative studies with normal cells.
A 4 kb region of the MMP-1 promoter DNA from a leukocyte genomic library was isolated and sequenced (Rutter et al. J. Cell Biochem. 66:322-336, 1997). DNA sequence analysis revealed that this clone contained only 1 G at position −1607 bp, resulting in the sequence 5′-AAGAT-3′ (SEQ ID NO: 1) (Rutter et al. J. Cell Biochem. 66:322-336, 1997). This sequence differs from that reported by others (Aho et al. Eur. J. Biochem. 247:503-510, 1997; Imai et al. Mol. Cell Biol. 14:7182-7194, 1994), wherein 2 Gs at that location which create the sequence 5′-AAGGAT-3′ (SEQ ID NO: 2) are described. The presence of 2 Gs at this site creates the sequence 5′-GGA-3′, which is a consensus sequence (Graves, J. B. Science 279:1000-1001, 1998) for a functional PEA3/EBS. This site has now been established to constitute a single nucleotide polymorphism (SNP). The full length DNA sequence of MMP-1 with only 1 G at position −1607 is depicted in SEQ ID NO: 3.
Based upon the effect of this SNP on the transcriptional activity, protein/DNA binding activity, and frequency of this SNP in normal fibroblasts and in melanoma tumor cells it has now been determined that detection of this SNP is useful in diagnosing and prognosticating cancer and other MMP-1 related diseases. It is also now believed that agents identified as inhibitors of binding of transcription factors to the Ets transcription factor binding site created by or resulting from this SNP will be useful in treating MMP-1 related diseases.